nrf2 truncation mutant construct δ2-86 amino acids #u617hel120-4 (GenScript corporation)

GenScript corporation nrf2 truncation mutant construct δ2-86 amino acids #u617hel120-4

a , Endogenous <t>NRF2</t> was IPed in MDA-MB-231 cells, and PtdIns(4,5)P 2 was analyzed by fluorescent IB. IgG was used as a negative control. Representative data from three independent experiments are shown. b , HEK293FT cells were transiently transfected with Flag-tagged NRF2 and processed for pulldown of PtdIns(4,5)P 2 48 h later. c , Analysis of protein-lipid interaction by MST with the dissociation constant (K d ) of each interaction indicated. d,e , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for IF against PtdIns(4,5)P 2 and NRF2. DAPI was used to stain the nuclei. The images ( d ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The graph ( e ) is shown as mean ± s.d. of three independent experiments with n=10 cells scored in each independent experiment. f, g , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for PLA between PtdIns(4,5)P 2 and NRF2. DAPI was used to stain nuclei. The images ( f ) were taken with a Leica SP8 confocal microscope. LASX (Leica) was used to quantify the PLA signal, and the graph ( g ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. Scale bar, 5 μm.

Nrf2 Truncation Mutant Construct δ2 86 Amino Acids #U617hel120 4, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nrf2 truncation mutant construct δ2-86 amino acids #u617hel120-4 - by Bioz Stars, 2026-09

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a , Endogenous NRF2 was IPed in MDA-MB-231 cells, and PtdIns(4,5)P 2 was analyzed by fluorescent IB. IgG was used as a negative control. Representative data from three independent experiments are shown. b , HEK293FT cells were transiently transfected with Flag-tagged NRF2 and processed for pulldown of PtdIns(4,5)P 2 48 h later. c , Analysis of protein-lipid interaction by MST with the dissociation constant (K d ) of each interaction indicated. d,e , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for IF against PtdIns(4,5)P 2 and NRF2. DAPI was used to stain the nuclei. The images ( d ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The graph ( e ) is shown as mean ± s.d. of three independent experiments with n=10 cells scored in each independent experiment. f, g , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for PLA between PtdIns(4,5)P 2 and NRF2. DAPI was used to stain nuclei. The images ( f ) were taken with a Leica SP8 confocal microscope. LASX (Leica) was used to quantify the PLA signal, and the graph ( g ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. Scale bar, 5 μm.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a , Endogenous NRF2 was IPed in MDA-MB-231 cells, and PtdIns(4,5)P 2 was analyzed by fluorescent IB. IgG was used as a negative control. Representative data from three independent experiments are shown. b , HEK293FT cells were transiently transfected with Flag-tagged NRF2 and processed for pulldown of PtdIns(4,5)P 2 48 h later. c , Analysis of protein-lipid interaction by MST with the dissociation constant (K d ) of each interaction indicated. d,e , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for IF against PtdIns(4,5)P 2 and NRF2. DAPI was used to stain the nuclei. The images ( d ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The graph ( e ) is shown as mean ± s.d. of three independent experiments with n=10 cells scored in each independent experiment. f, g , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for PLA between PtdIns(4,5)P 2 and NRF2. DAPI was used to stain nuclei. The images ( f ) were taken with a Leica SP8 confocal microscope. LASX (Leica) was used to quantify the PLA signal, and the graph ( g ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. Scale bar, 5 μm.

Article Snippet: The full-length NRF2 construct (#OHU26812) and each of the NRF2 truncation mutant constructs Δ2-86 amino acids (# U617HEL120-4), Δ87-204 amino acids (#U617HEL120-5), Δ205-400 amino acids (# U617HEL120-6), and Δ401-605 amino acids (#U617HEL120-7) were purchased from GenScript.

Techniques: Negative Control, Transfection, Staining, Microscopy

a , Representative MST curves for the interaction between protein NRF2 and different phosphoinositide ligands at different concentrations. b, Representative IB of NRF2 expression in MDA-MB-231 cells treated with different stressors for 4 h: Cisplatin (30μM), tBHQ (100uM), and sulforaphane (30μM), LCS-1 (10μM) and Diethyl maleate (DEM 200μM). c , MDA-MB-231 cells were treated with 200 μM DEM for 0-4 h, and NRF2 expression was analyzed by IB.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a , Representative MST curves for the interaction between protein NRF2 and different phosphoinositide ligands at different concentrations. b, Representative IB of NRF2 expression in MDA-MB-231 cells treated with different stressors for 4 h: Cisplatin (30μM), tBHQ (100uM), and sulforaphane (30μM), LCS-1 (10μM) and Diethyl maleate (DEM 200μM). c , MDA-MB-231 cells were treated with 200 μM DEM for 0-4 h, and NRF2 expression was analyzed by IB.

Techniques: Expressing

a,b , The expression of NRF2 was analyzed by IB in MDA-MB-231 cells with deletion of PIPKIα, PIPKIγ, PIPKIIα, or PIPKIIβ ( a ). The intensity of the immunoblots was quantified, and the graph is shown ( b ) as mean ± s.d of n=3 independent experiments. c , PIPKIγ was stably knocked down (KD) by two shRNAs in HCT116 cells. Cells were treated with vehicle or 100 μM tBHQ for 4 h. The expression of NRF2 was analyzed by IB, and the graph is shown as mean ± s.d of n=3 independent experiments. d , MDA-MB-231 cells (parental or with PIPKIγ deletion) were treated with 100 μM tBHQ for 4h. NRF2 protein expression was analyzed by IB, e , MDA-MB-231 cells transfected with control siRNAs or siRNAs targeting PIPKIγ were treated with or without 10 μM MG132 for the indicated time. NRF2 protein levels were determined by IB.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a,b , The expression of NRF2 was analyzed by IB in MDA-MB-231 cells with deletion of PIPKIα, PIPKIγ, PIPKIIα, or PIPKIIβ ( a ). The intensity of the immunoblots was quantified, and the graph is shown ( b ) as mean ± s.d of n=3 independent experiments. c , PIPKIγ was stably knocked down (KD) by two shRNAs in HCT116 cells. Cells were treated with vehicle or 100 μM tBHQ for 4 h. The expression of NRF2 was analyzed by IB, and the graph is shown as mean ± s.d of n=3 independent experiments. d , MDA-MB-231 cells (parental or with PIPKIγ deletion) were treated with 100 μM tBHQ for 4h. NRF2 protein expression was analyzed by IB, e , MDA-MB-231 cells transfected with control siRNAs or siRNAs targeting PIPKIγ were treated with or without 10 μM MG132 for the indicated time. NRF2 protein levels were determined by IB.

Techniques: Expressing, Western Blot, Stable Transfection, Transfection, Control

a , b , MDA-MB-231 cells were transfected with control siRNAs or siRNAs targeting PIPKIγ, and NRF2 was analyzed by IB 72 h later. NRF2 IB was quantified, and the graph is shown as mean ± s.d of n=3 independent experiments. c , MDA-MB-231 cells were transfected with control siRNAs or siRNAs targeting PIPKIγ for 72 h, and NFE2L2 mRNA levels were determined by RT-PCR. NFE2L2 mRNA levels were normalized to GAPDH mRNA. The graph is shown as mean ± s.d of n=3 independent experiments.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a , b , MDA-MB-231 cells were transfected with control siRNAs or siRNAs targeting PIPKIγ, and NRF2 was analyzed by IB 72 h later. NRF2 IB was quantified, and the graph is shown as mean ± s.d of n=3 independent experiments. c , MDA-MB-231 cells were transfected with control siRNAs or siRNAs targeting PIPKIγ for 72 h, and NFE2L2 mRNA levels were determined by RT-PCR. NFE2L2 mRNA levels were normalized to GAPDH mRNA. The graph is shown as mean ± s.d of n=3 independent experiments.

Techniques: Transfection, Control, Reverse Transcription Polymerase Chain Reaction

a , b , MDA-MB-231 ( a ) or HCT116 cells ( b ) cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous PIPKIγ was IPed, and NRF2 was analyzed by IB. Representative IBs of three independent experiments are shown. c,d , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for IF against PIPKIγ and NRF2. DAPI was used to stain nuclei. The images ( c ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The graph ( d ) is shown as mean ± s.d of three independent experiments with n=10 cells scored in each independent experiment. e,f , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for PLA between PIPKIγ and NRF2. DAPI was used to stain nucleic acids. The images ( e ) were taken with a Leica SP8 confocal microscope. The red PLA signal was quantified by LASX (Leica), and the graph ( f ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. g,h , MDA-MB-231 cells with or without PIPKIγ KD were treated with vehicle or 200 μM DEM for 4 h before being processed for PLA between PtdIns(4,5)P 2 and NRF2 ( g ). DAPI was used to stain nucleic acids. The red PLA signal was quantified by LAS X (Leica). The experiments were repeated three times. The graph is shown as mean ± s.d. of n=15 cells from one representative experiment ( h ). Scale bar,5 μm.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a , b , MDA-MB-231 ( a ) or HCT116 cells ( b ) cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous PIPKIγ was IPed, and NRF2 was analyzed by IB. Representative IBs of three independent experiments are shown. c,d , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for IF against PIPKIγ and NRF2. DAPI was used to stain nuclei. The images ( c ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The graph ( d ) is shown as mean ± s.d of three independent experiments with n=10 cells scored in each independent experiment. e,f , MDA-MB-231 cells were treated with 200 μM DEM for 4 h before being processed for PLA between PIPKIγ and NRF2. DAPI was used to stain nucleic acids. The images ( e ) were taken with a Leica SP8 confocal microscope. The red PLA signal was quantified by LASX (Leica), and the graph ( f ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. g,h , MDA-MB-231 cells with or without PIPKIγ KD were treated with vehicle or 200 μM DEM for 4 h before being processed for PLA between PtdIns(4,5)P 2 and NRF2 ( g ). DAPI was used to stain nucleic acids. The red PLA signal was quantified by LAS X (Leica). The experiments were repeated three times. The graph is shown as mean ± s.d. of n=15 cells from one representative experiment ( h ). Scale bar,5 μm.

Techniques: Staining, Microscopy

a,b , Endogenous NRF2 was IPed from MDA-MB-231 ( a ) or MCF7 ( b ) cells, and HSP27, αB-crystallin, HSP40, HSP60, and HSP90 were analyzed by IB. IgG was used as a negative control. Representative data from three independent experiments are shown. c , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4h. Endogenous NRF2 was IPed, and HSP27 was analyzed by IB. Represent data from three independent experiments are shown. d , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous HSP27 was IPed, and NRF2 was analyzed by IB. Represent data from three independent experiments are shown. e,f , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for IF staining against HSP27 and NRF2 ( e ). DAPI was used to stain nuclei. The images ( e ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The experiments were repeated three times, and graph ( f ) is shown as mean ± s.d of three independent experiments with n=10 cells scored in each independent experiment. g,h, MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for PLA between HSP27 and NRF2. DAPI was used to stain nuclei. The images ( g ) were taken with a Leica SP8 confocal microscope. The red PLA signal was quantified by LASX (Leica), and the graph ( h ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. Scale bar, 5 μm.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a,b , Endogenous NRF2 was IPed from MDA-MB-231 ( a ) or MCF7 ( b ) cells, and HSP27, αB-crystallin, HSP40, HSP60, and HSP90 were analyzed by IB. IgG was used as a negative control. Representative data from three independent experiments are shown. c , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4h. Endogenous NRF2 was IPed, and HSP27 was analyzed by IB. Represent data from three independent experiments are shown. d , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous HSP27 was IPed, and NRF2 was analyzed by IB. Represent data from three independent experiments are shown. e,f , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for IF staining against HSP27 and NRF2 ( e ). DAPI was used to stain nuclei. The images ( e ) were taken with a Leica SP8 confocal microscope and processed by ImageJ. The experiments were repeated three times, and graph ( f ) is shown as mean ± s.d of three independent experiments with n=10 cells scored in each independent experiment. g,h, MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h before being processed for PLA between HSP27 and NRF2. DAPI was used to stain nuclei. The images ( g ) were taken with a Leica SP8 confocal microscope. The red PLA signal was quantified by LASX (Leica), and the graph ( h ) is shown as mean ± s.d of three independent experiments with n=15 cells scored in each independent experiment. Scale bar, 5 μm.

Techniques: Negative Control, Staining, Microscopy

a,b, Recombinant His-tagged NRF2 immobilized on NRF2 beads was incubated with the indicated amount of HSP27 ( a ) or αB-crystallin ( b ) protein in vitro . The complex was pulled down, and sHSP-bound NRF2 was analyzed by IB. The graph is shown as mean ± s.d of n=3 independent experiments. AU is an arbitrary unit. c,d , Representative MST curves for the interaction between protein NRF2 and HSP27 ( c ) or αB-crystallin ( d ) at different concentrations. e , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous NRF2 was then IPed, and αB-crystallin was analyzed by immunoblotting. Represent data from three independent experiments are shown. f, MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4h before being processed for PLA between αB-crystallin and NRF2. DAPI was used to stain nucleic acids.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a,b, Recombinant His-tagged NRF2 immobilized on NRF2 beads was incubated with the indicated amount of HSP27 ( a ) or αB-crystallin ( b ) protein in vitro . The complex was pulled down, and sHSP-bound NRF2 was analyzed by IB. The graph is shown as mean ± s.d of n=3 independent experiments. AU is an arbitrary unit. c,d , Representative MST curves for the interaction between protein NRF2 and HSP27 ( c ) or αB-crystallin ( d ) at different concentrations. e , MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4 h. Endogenous NRF2 was then IPed, and αB-crystallin was analyzed by immunoblotting. Represent data from three independent experiments are shown. f, MDA-MB-231 cells were treated with vehicle or 200 μM DEM for 4h before being processed for PLA between αB-crystallin and NRF2. DAPI was used to stain nucleic acids.

Techniques: Recombinant, Incubation, In Vitro, Western Blot, Staining

a,b MDA-MB-468 cells were transiently transfected with control siRNAs or siRNAs targeting αB-crystallin ( a ) or HSP27 ( b ) for 72 h. Vehicle or 100 μM tHBQ was added for the final 4 h, and NRF2 expression was analyzed by IB.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a,b MDA-MB-468 cells were transiently transfected with control siRNAs or siRNAs targeting αB-crystallin ( a ) or HSP27 ( b ) for 72 h. Vehicle or 100 μM tHBQ was added for the final 4 h, and NRF2 expression was analyzed by IB.

Techniques: Transfection, Control, Expressing

a , Schematic representation of NRF2 truncation mutants used in the study. b, Recombinant His-Tag NRF2 truncation mutants were immobilized on HSP27 or αB-crystallin beads, and HSP27 or αB-crystallin bound to each mutant was analyzed by IB. Representative data from three independent experiments are shown. c , Recombinant His-Tag NRF2 truncation mutants were pulled down by PtdIns(4,5)P 2 beads, and PtdIns(4,5)P 2 bound to each NRF2 mutant was analyzed by IB. Representative data from three independent experiments are shown. d , e , 6 nM recombinant His-tagged NRF2 and 200 nM HSP27 or 120 nM αB-crystallin were incubated with the indicated concentrations of PtdIns(4,5)P 2 . NRF2 was pulled down, and the bound HSP27 (d) or αB-crystallin (e) was analyzed by IB. Representative data from three independent experiments are shown.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a , Schematic representation of NRF2 truncation mutants used in the study. b, Recombinant His-Tag NRF2 truncation mutants were immobilized on HSP27 or αB-crystallin beads, and HSP27 or αB-crystallin bound to each mutant was analyzed by IB. Representative data from three independent experiments are shown. c , Recombinant His-Tag NRF2 truncation mutants were pulled down by PtdIns(4,5)P 2 beads, and PtdIns(4,5)P 2 bound to each NRF2 mutant was analyzed by IB. Representative data from three independent experiments are shown. d , e , 6 nM recombinant His-tagged NRF2 and 200 nM HSP27 or 120 nM αB-crystallin were incubated with the indicated concentrations of PtdIns(4,5)P 2 . NRF2 was pulled down, and the bound HSP27 (d) or αB-crystallin (e) was analyzed by IB. Representative data from three independent experiments are shown.

Techniques: Recombinant, Mutagenesis, Incubation

a, MDA-MB-231 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27, and NRF2 expression was analyzed by IB 72 h later. The graph is shown as mean ± s.d of n=3 independent experiments. b , RT-PCR analysis of NFE2L2 mRNA in MDA-MB-231 cells. NFE2L2 mRNA levels were normalized to GAPDH mRNA. The graph is shown as mean ± s.d of n=4 independent experiments. c , MDA-MB-231 cells transfected with control siRNAs or siRNAs targeting HSP27 were treated with or without 10 μM MG132 for the indicated time. NRF2 protein levels were determined by IB. d,e, HCT116 ( d ) or HS578 ( e ) cells were transiently transfected with control siRNAs or siRNAs targeting HSP27. 68 h later, transfected cells were treated with vehicle or 100 μM tBHQ for an additional 4 h, and NRF2 expression was then analyzed by IB. The graph is shown as mean ± s.d of n=3 independent experiments. f, A549 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27, and NRF2 expression was analyzed by IB 72 h later. The graph is shown as mean ± s.d of n=3 independent experiments. g , 0.5μg of recombinant His-tagged NRF2 and 9 μg KEAP1 were incubated with the indicated concentrations of HSP27. NRF2 was pulled down, and the bound KEAP1 and HSP27 were analyzed by IB.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a, MDA-MB-231 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27, and NRF2 expression was analyzed by IB 72 h later. The graph is shown as mean ± s.d of n=3 independent experiments. b , RT-PCR analysis of NFE2L2 mRNA in MDA-MB-231 cells. NFE2L2 mRNA levels were normalized to GAPDH mRNA. The graph is shown as mean ± s.d of n=4 independent experiments. c , MDA-MB-231 cells transfected with control siRNAs or siRNAs targeting HSP27 were treated with or without 10 μM MG132 for the indicated time. NRF2 protein levels were determined by IB. d,e, HCT116 ( d ) or HS578 ( e ) cells were transiently transfected with control siRNAs or siRNAs targeting HSP27. 68 h later, transfected cells were treated with vehicle or 100 μM tBHQ for an additional 4 h, and NRF2 expression was then analyzed by IB. The graph is shown as mean ± s.d of n=3 independent experiments. f, A549 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27, and NRF2 expression was analyzed by IB 72 h later. The graph is shown as mean ± s.d of n=3 independent experiments. g , 0.5μg of recombinant His-tagged NRF2 and 9 μg KEAP1 were incubated with the indicated concentrations of HSP27. NRF2 was pulled down, and the bound KEAP1 and HSP27 were analyzed by IB.

Techniques: Transfection, Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Recombinant, Incubation

a, HCT116 cells expressing control shRNA or shRNAs targeting PIPKIγ were treated with 200 μM DEM for 4 h. The expression of PIPKIγ, HO-1, and NRF2 were analyzed by IB. Representative data from three independent experiments are shown. b, MDA-MB-231 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27 for 72 h. Cells were then treated with vehicle or 200 μM DEM for an additional 4 h, and HO-1, NRF2, and HSP27 expression was analyzed by IB. Representative data from three independent experiments are shown. c,d, ROS levels were determined in MDA-MB-231 and HCT116 cells transiently transfected with control siRNAs or siRNAs targeting HSP27 ( c ) or PIPKIγ ( d ) for 72 h. Cells were treated with vehicle or 200 μM DEM for the final 4 h before analyzing ROS levels. The graph is shown as mean ± s.d of n=3 independent experiments. e,f , Cell viability was determined by MTT assay in MDA-MB-231 and HCT116 cells transiently transfected with control siRNAs or siRNAs targeting HSP27 ( e ) or PIPKIγ ( f ) for 72 h. Cells were treated with vehicle or 200 μM DEM for the final 4 h before analyzing cell viability. The graph is shown as mean ± s.d of n=3 independent experiments. g , A schematic model of NRF2 regulation by PIPKIγ, PtdIns(4,5)P 2 and sHSPs.

Journal: bioRxiv

Article Title: Regulation of NRF2 by Phosphoinositides and Small Heat Shock Proteins

doi: 10.1101/2023.10.26.564194

Figure Lengend Snippet: a, HCT116 cells expressing control shRNA or shRNAs targeting PIPKIγ were treated with 200 μM DEM for 4 h. The expression of PIPKIγ, HO-1, and NRF2 were analyzed by IB. Representative data from three independent experiments are shown. b, MDA-MB-231 cells were transiently transfected with control siRNAs or siRNAs targeting HSP27 for 72 h. Cells were then treated with vehicle or 200 μM DEM for an additional 4 h, and HO-1, NRF2, and HSP27 expression was analyzed by IB. Representative data from three independent experiments are shown. c,d, ROS levels were determined in MDA-MB-231 and HCT116 cells transiently transfected with control siRNAs or siRNAs targeting HSP27 ( c ) or PIPKIγ ( d ) for 72 h. Cells were treated with vehicle or 200 μM DEM for the final 4 h before analyzing ROS levels. The graph is shown as mean ± s.d of n=3 independent experiments. e,f , Cell viability was determined by MTT assay in MDA-MB-231 and HCT116 cells transiently transfected with control siRNAs or siRNAs targeting HSP27 ( e ) or PIPKIγ ( f ) for 72 h. Cells were treated with vehicle or 200 μM DEM for the final 4 h before analyzing cell viability. The graph is shown as mean ± s.d of n=3 independent experiments. g , A schematic model of NRF2 regulation by PIPKIγ, PtdIns(4,5)P 2 and sHSPs.

Techniques: Expressing, Control, shRNA, Transfection, MTT Assay

nrf2 truncation mutant construct δ205 to 400 amino acids Search Results

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